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The escalating presence of microplastics and heavy metals in marine environments significantly jeopardizes ecological stability and human health. Despite this, research on the combined effects of microplastics/nanoplastics (MPs/NPs) and heavy metals on marine organisms remains limited. This study evaluated the impact of two sizes of polystyrene beads (approximately 2 µm and 200 nm) combined with cadmium (Cd) on the ciliate species Euplotes vannus. Results demonstrated that co-exposure of MPs/NPs and Cd markedly elevated reactive oxygen species (ROS) levels in ciliates while impairing antioxidant enzyme activities, thus enhancing oxidative damage and significantly reducing carbon biomass in ciliates. Transcriptomic profiling indicated that co-exposure of MPs/NPs and Cd potentially caused severe DNA damage and protein oxidation, as evidenced by numerous differentially expressed genes (DEGs) associated with mismatch repair, DNA replication, and proteasome function. Integrated transcriptomic and metabolomic analysis revealed that DEGs and differentially accumulated metabolites (DAMs) were significantly enriched in the TCA cycle, glycolysis, tryptophan metabolism, and glutathione metabolism. This suggests that co-exposure of MPs/NPs and Cd may reduce ciliate abundance and carbon biomass by inhibiting energy metabolism and antioxidant pathways. Additionally, compared to MPs, the co-exposure of NPs and Cd exhibited more severe negative effects due to the larger specific surface area of NPs, which can carry more Cd. These findings provide novel insights into the toxic effects of MPs/NPs and heavy metals on protozoan ciliates, offering foundational data for assessing the ecological risks of heavy metals exacerbated by MPs/NPs.

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The impacts and toxicological mechanisms of microplastics (MPs) or heavy metals on aquatic ecosystems have been the subject of extensive research and initial understanding. However, the combined toxicity of co-pollutants on organisms and cumulative toxic effects along the food chain are still underexplored. In this study, the ciliate protozoan Paramecium caudatum and zebrafish Danio rerio were used to represent the microbial loop and the higher trophic level, respectively, to illustrate the progressive exposure of MPs and cadmium (Cd2+). The findings indicate that MPs (ca. 1 ×105 items/L) containing with Cd2+ (below 0.1 µg/L) could permeate the bodies of zebrafish through trophic levels after primary ingestion by ciliates. This could cause adverse effects on zebrafish, including alterations in bioindicators (total sugar, triglycerides, lactate, and glycogen) associated with metabolism, delayed hepatic development, disruption of intestinal microbiota, DNA damage, inflammatory responses, and abnormal cellular apoptosis. In addition, the potential risks associated with the transfer of composite pollutants through the microbial loop into traditional food chain were examined, offering novel insights on the evaluation of the ecological risks associated with MPs. As observed, understanding the bioaccumulation and toxic effects of combined pollutants in zebrafish holds crucial implications for food safety and human health.

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Protozoan ciliates represent a common biological contaminant during microalgae cultivation, which will lead to a decline in microalgae productivity. This study investigated the effectiveness of sodium dodecyl benzene sulfonate (SDBS) in controlling ciliate populations within microalgae cultures. SDBS concentrations of 160 mg/L and 100 mg/L were found to effectively manage the representative species of ciliates contamination by Euplotes vannus and Uronema marinum during the cultivation of Synechococcus and Chlorella, and the growth vitality of microalgae has been restored. Additionally, SDBS at these concentrations reduced oxidative stress resistance and induced membrane damage to remove biological pollutants by modulating enzyme activity, affecting lipid, energy, amino acid metabolism pathways, and processes such as translation and protein folding. This research provides insights into the mechanisms through which SDBS effectively combats protozoan ciliates during the microalgal cultivation. This contributes to reduce biological pollution, ensure the overall productivity and healthy and sustainable management of microalgae ecosystems.

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Micro/nano-plastics (MPs/NPs) represent an emerging contaminant, posing a significant threat to oceanic halobios. While the adverse effects of joint pollutants on marine organisms are well-documented, the potential biological impacts on the food chain transmission resulting from combinations of MPs/NPs and heavy metals (HMs) remain largely unexplored. This study exposed the microbial loop to combined contaminants (MPs/NPs + HMs) for 48h, bacteria and contaminants are washed away before feeding to the traditional food chain, employing microscopic observation, biochemical detection, and transcriptome analysis to elucidate the toxicological mechanisms of the top predator. The findings revealed that MPs/NPs combined with Cd2+ could traverse both the microbial loop and classical food chain. Acute exposure significantly affected the carbon biomass of the top predator Tigriopus japonicus (75.8% lower). Elevated antioxidant enzyme activity led to lipid peroxidation, manifesting in increased malondialdehyde levels. Transcriptome sequencing showed substantial differential gene expression levels in T. japonicus under various treatments. The upregulation of genes associated with apoptosis and inflammatory responses, highlighting the impact of co-exposure on oxidative damage and necroptosis within cells. Notably, NPs-Cd exhibited stronger toxicity than MPs-Cd. NPs-Cd led to a greater decrease in the biomass of top predators, accompanied by lower activities of GSH, SOD, CAT, and GSH-PX, resulting in increased production of lipid peroxidation product MDA and higher oxidative stress levels. This investigation provides novel insights into the potential threats of MPs/NPs combined with Cd2+ on the microbial loop across traditional food chain, contributing to a more comprehensive assessment of the ecological risks associated with micro/nano-plastics and heavy metals.

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Electrochemical advanced oxidation processes (EAOPs) have shown great potential for the treatment of shale gas produced water (SGPW). In this study, we investigated the transformation of dissolved organic matter (DOM) during EAOPs of SGPW and the formation of toxic halogenated by-products at various current densities, using fluorescence spectroscopy and Fourier transform ion cyclotron resonance mass spectrometry. We found that the priority of DOM removal was terrestrial humic-like > microbial humic-like > protein-like substances. Non-Halogenated organic compounds (non-HOCs) and HOCs were predominantly CHO, and CHOCl/CHOBr compounds in EAOP-treated SGPW, respectively. As applied current density and treatment time increased, the production of oxyhalides increased, with chlorate > bromate > perchlorate. Meanwhile, most DOM was mineralized, resulting in residual products with higher modified aromaticity index (AImod) and nominal oxidation state of carbon (NOSC). The resistants had lower mass-to-charge ratio (m/z), AImod, NOSC, and double bond equivalent minus oxygen per carbon ((DBE-O)/C). The dominant reactions were the addition of tri-oxygen and deallyl. Bromine addition dominated the reactions of halogenating addition, while chlorine addition took second place. Furthermore, the acute toxicity of SGPW was positively correlated with inorganic halogenated by-products. This study contributes to the understanding and improvement of EAOPs for the treatment of SGPW.

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Produced water (PW) is the largest waste stream generated by hydraulic fracturing in an unconventional shale gas reservoir. Oxidation processes (OPs) are frequently used as advanced treatment method in highly complicated water matrix treatments. However, the degradation efficiency is the main focus of research, organic compounds and their toxicity have not been properly explored. Here, we obtained the characterization and transformation of dissolved organic matters of PW samples from the first shale gas field of China by two selected OPs using FT-ICR MS. CHO, CHON, CHOS, and CHONS heterocyclic compounds associated with lignins/CRAM-like, aliphatic/proteins, and carbohydrates compounds were the major organic compounds identified. Electrochemical Fe2+/HClO oxidation preferentially removed aromatic structures, unsaturated hydrocarbons, and tannin compounds with a double-bond equivalence (DBE) value below 7 to more saturated compounds. Nevertheless, Fe (VI) degradation manifested in CHOS compounds with low DBE values, especially single bond compounds. Oxygen- and Sulfur-containing substances, primarily O4-11, S1O3-S1O12, N1S1O4, and N2S1O10 classes, were the main recalcitrant components in OPs. The toxicity assessment showed that the free-radical-formed Fe2+/HClO oxidation could cause significant DNA damage. Therefore, the toxicity response byproducts need spcial attention when conducting OPs. Our results led to discussions on designing appropriate treatment strategies and the development of PW discharge or reuse standards.

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Electrochemical advanced oxidation process (EAOP) is recommended for high-strength refractory organics wastewater treatment, but the accompanying chlorinated byproduct generation becomes a bottleneck that limits the application of this technology to actual wastewater. In this study, we applied EAOP (0.4-40 mA cm-2) to treat ultrafiltration effluent of an actual landfill leachate, and quantitatively assessed the toxicities of the dominant chlorinated byproducts in EAOP-treated effluent. Considering both toxic effect and dose, it followed the order: active chlorine > chlorate > perchlorate > organochlorines. The toxic active chlorine could spontaneously decompose by settling. And secondary bioreactor originally serving for denitrification could be used to reduce perchlorate and chlorate. The effects of residual active chlorine and extra carbon addition on simultaneous denitrification, perchlorate, and chlorate reduction were investigated. It seemed that 20 mg of active chlorine was an acceptable level to bioactivity, and sufficient electron donors favored the removal of chlorate and perchlorate. Pseudomonas was identified as an active chlorine tolerant chlorate-reducing bacteria. And Thauera was responsible for perchlorate reduction under the conditions of sufficient carbon source supply. Our results confirmed that the perchlorate and chlorate concentrations in the effluent below their health advisory levels were achievable, solving the issue of toxic chlorinated byproduct generation during EAOP. This study provided a solution to realistic application of EAOP to treat high chloride wastewater.

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The waste product phosphogypsum (PG) is produced in phosphoric acid production processes. Its storage requires large amounts of land resources and poses serious environmental risks. In this work, detailed experimental research was carried out to investigate the potential reuse of PG after calcination modification as a novel building material for cast-in-place concrete products. The calcination modification mechanism was studied, and the environmental risk assessment of modified PG was presented. The results showed that the calcination modification includes crystal phase transformation, removal of impurities, and modifying the pH value. The calcination was carried out at 280 °C for 5 h, where the resulting product was a pH value of 7.1, and the soluble fluorine and phosphorus removal rates reached up to 69.2% and 71.2%, respectively. These removal rates met the requirements of the China national standard Phosphogypsum (GB/T 23456-2018). To ensure the environmental safety, ecological risk assessment methods for determining the leaching toxicity of the modified PG were employed. The toxicity of Ba and P elements in the modified PG products was assessed, as well as the leaching toxicity concentrations of all particular heavy metals, which were found well below the limits set by the national standards. All the results presented strongly suggest that the 280 °C modified PG presented here has excellent application potential as a raw component in building materials.

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The vegetation deterioration and pollution expansion from non-ferrous metal tailings pond have been found in many countries leading to water soil erosion and human health risk. Conventional ecological remediation technologies of mine tailings such as capping were costly and elusive. This study provided an economic and effective model as an alternative by substrate amelioration and vegetation restoration. A field experiment was carried out on a silver tailings pond in southwest China. Tailings substrate was ameliorated by adding organic matter (decomposed chicken manure, DCM), structural conditioner (polyacrylamide, PAM), water-retaining agent (acrylic acid-bentonite water-retaining agent, AAB), and heavy metal immobilizer (biofuel ash, BFA), which were optimized by laboratory experiment. Native heavy metal hyperaccumulator, Bidens pilosa, was colonized. Vegetation coverage and plant height of Bidens pilosa reached about 80% and over 30 cm respectively after 3 months, and the turbidity of tailings leaching solution decreased by 60%. The practice showed that the proportion of available heavy metals in tailings substrate was significantly lower than that in the soil surrounding mining area. Immobilization didn't have stabilization effect on Cd, Zn, and Pb, and As was only 0.002%, phytoremediation had stabilization effect of Cd, Zn, As, and Pb were 2.5-3.5%, 1-2%, 0.25-0.5%, and 0.25-0.75%. Phytoremediation was more effective significantly in controlling heavy metal pollution risk of tailings than immobilization. These results provided a new ecological remediation OSA-NHC model, meaning a combination of optimal substrate amelioration and native hyperaccumulator colonization, which could achieve vegetation restoration and augment heavy metal pollution control in non-ferrous metal tailings pond.

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The unique ability of Anammox bacteria to metabolize short-chain fatty acids have been demonstrated. However, the potential contributions of active Anammox species to carbon utilization in a mixotrophic Anammox-denitrification process are less well understood. In this study, we combined genome-resolved metagenomics and DNA stable isotope probing (DNA-SIP) to characterize an Anammox process fed with acetate under COD/TN ratios of around 0.30-0.40 and low nitrogen-loading rates. A draft genome of "Candidatus Jettenia caeni" and a novel species that was phylogenetically close to "Candidatus Brocadia sinica" were recovered. Essential genes encoding the key enzymes for acetate metabolism and dissimilatory nitrate reduction to ammonium were identified in the two Anammox draft genomes. The DNA-SIP revealed that Ignavibacterium, "Candidatus Jettenia caeni," Thauera, Denitratisoma, and Calorithrix predominantly contributed to organic carbon utilization in the acetate-fed Anammox process. In particular, the "Candidatus Jettenia caeni" accounted for a higher proportion of 13C-DNA communities than "Candidatus Brocadia sinica." This result well confirmed the theory of maintenance energy between the interspecies competition of the two Anammox species under low nitrogen-loading rates. Our study revealed its potential important role of the Anammox genus "Candidatus Jettenia" in the treatment of wastewater containing low organic matter and ammonia.

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Shale gas-produced water (PW), the waste fluid generated during gas production, contains a large number of organic contaminants and high salinity matrix. Previous studies generally focused on the end-of-pipe treatment of the PW and ignored the early collection process. In this study, the transformation of the molecular composition and microbial community structure of the PW in the transportation and storage process (i.e., from the gas-liquid separator to the storage tank) were investigated. As the PW was transported from the gas-liquid separator to the portable storage tank, the dissolved organic matter (DOM) showed greater saturation, less oxidation, and lower polarity. DOMs with high O/C and low H/C ratios (numbers of oxygen and hydrogen divided by numbers of carbon) were eliminated, which may be due to precipitation or adsorption by the solids suspended in the PW. The values of double-bond equivalent (DBE), DBE/C (DBE divided by the number of carbon), and aromatic index (AI) decreased, likely because of the microbial degradation of aromatic compounds. The PW in the gas-liquid separator presented a lower biodiversity than that in the storage tank. The microbial community in the storage tank showed the coexistence of anaerobes and aerobes. Genera related to biocorrosion and souring were detected in the two facilities, thus indicating the necessity of more efficient anticorrosion strategies. This study helps to enhance the understanding of the environmental behavior of PW during shale gas collection and provides a scientific reference for the design and formulation of efficient transportation and storage strategies to prevent and control the environmental risk of shale gas-derived PW.

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Obligate aerobic methanotrophs have been proven to oxidize methane and participate in denitrification under hypoxic conditions. However, this phenomenon and its metabolic mechanism have not been investigated in detail in aerobic methane oxidation coupled to denitrification (AME-D) process. In this study, a type of hypoxic AME-D consortium was enriched and operated for a long time in a CH4-cycling bioreactor with strict anaerobic control and the nitrite removal rate reached approximately 50 mg N/L/d. Metagenomics combined with DNA stable-isotope probing demonstrated that the genus Methylomonas, which constitutes type I aerobic methanotrophs, was the dominant member and contributed to methane oxidation and partial denitrification. Metagenomic binning recovered a near-complete (98%) draft genome affiliated with the family Methylococcaceae containing essential genes that encode nitrite reductase (nirK), nitric oxide reductase (norBC) and hydroxylamine dehydrogenase (hao). Metabolic reconstruction of the selected Methylococcaceae genomes also revealed a potential link between methanotrophy and partial denitrification.

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Tannery sludge usually has high content of trivalent chromium (Cr(III)) and ammonium-nitrogen (NH4+-N). It is important to make a critical evaluation of the releasing behaviors of Cr(III) and NH4+-N from tannery sludge before its use on improving soil fertility in agricultural applications. For this purpose, static batch and dynamic leaching experiments with different mathematical models were carried out to simulate the Cr(III) and NH4+-N releasing kinetics from tannery sludge sampled in a typical tannery disposal site in North China, and their influencing factors were also discussed. The results showed that a larger solid-liquid ratio, a higher temperature, and a lower pH value of the leaching solution were beneficial for the release of Cr(III) and NH4+-N from the tannery sludge. The release kinetics of Cr(III) and NH4+-N followed parabolic diffusion and simple Elovich models both in the static and dynamic leaching conditions, indicating that the release was a complex heterogeneous diffusion process. The NH4+-N was easy to be leached out and its released amount reached 3.14 mg/g under the dynamic leaching condition (pH 7), whereas the released amount of the Cr(III) was only 0.27 µg/g from the tannery sludge. There was a positive correlation coefficient between dissolved Fe and Cr(III) in the leachate under different leaching conditions, and the calculated average ratio of Fe/Cr(III) concentration was 3.56, indicating that the small amount of the released Cr(III) came from the dissolution of Cr0.25Fe0.75(OH)3 minerals in tannery sludge.

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Responses of a microbial community in the completely autotrophic nitrogen removal over nitrite (CANON) process, which was shocked by a pH of 11.0 for 12 h, were investigated. During the recovery phase, the performance, anaerobic ammonia oxidation (anammox) activity, microbial community, and correlation of bacteria as well as the influencing factors were evaluated synchronously. The performance of the CANON process deteriorated rapidly with a nitrogen removal rate (NRR) of 0.13 kg·m-3·d-1, and Firmicutes, spore-forming bacteria, were the dominant phyla after alkaline shock. However, it could self-restore within 107 days after undergoing four stages, at which Planctomycetes became dominant with a relative abundance of 64.62%. Network analysis showed that anammox bacteria (Candidatus Jettenia, Kuenenia, and Brocadia) were positively related to some functional bacteria such as Nitrosomonas, SM1A02, and Calorithrix. Canonical correspondence analysis presented a strong correlation between the microbial community and influencing factors during the recovery phase. With the increase of nitrogen loading rate, the decrease of free nitrous acid and the synergistic effects, heme c content, specific anammox activity (SAA), NRR, and the abundance of dominant genus increased correspondingly. The increase of heme c content regulates the quorum sensing system, promotes the secretion of extracellular polymeric substances, and further improves SAA, NRR, and the relative abundance of the dominant genus. This study highlights some implications for the recovery of the CANON reactor after being exposed to an alkaline shock.

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The degradation of roxarsone, an extensively used organoarsenic feed additive, occurs quickly under anaerobic conditions with microorganisms playing an important role in its degradation. Here, an anaerobic bacterial consortium that effectively degraded roxarsone was isolated, and its degradation efficiency and community changes along a roxarsone concentration gradient under anaerobic conditions were assessed. We used batch experiments to determine the roxarsone degradation rates, as well as the bacterial community structure and diversity, at initial roxarsone concentrations of 50, 100, 200, and 400 mg/kg. The results showed that roxarsone was degraded completely within 28, 28, 36, and 44 hr at concentrations of 50, 100, 200, and 400 mg/kg, respectively. The anaerobic bacterial consortium displayed considerable potential to degrade roxarsone, as the degradation rate increased with increasing roxarsone concentrations. Roxarsone promoted microbial growth, and in turn, the microorganisms degraded the organoarsenic compound, with the functional bacterial community varying between different roxarsone concentrations. Lysinibacillus, Alkaliphilus, and Proteiniclasticum were the main genera composing the roxarsone-degrading bacterial community.

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Anaerobic ammonium oxidation (anammox) pathway is sensitive to organic matter, and its recovery requires reliable evidence regarding the dominance of anammox in N-removal. This study showed that the anammox process deteriorated, with N-removal efficiencies rapidly decreasing from 87.2 to 45.7% when reactors were exposed to COD shocks of 1.12, 2.24 and 3.36 g L-1 (COD/N ratio 2, 4 and 6). Comprehensive assessments of water quality, microbial characteristics and isotope analysis were adopted to investigate anammox recovery. Operational performance took 8-20 days to recover; anammox relative abundance recovered after 20 days, based on the results of fluorescence in situ hybridisation and quantitative PCR; and the anammox pathway contributed to 80.0-91.5% of N-loss 40 days after COD shock terminated, based on the results of the isotope labelling experiment. Therefore, a complete recovery required 40 days. The isotope labelling method supplied a reliable reference for recovery assessment of anammox system in real-world applications.

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The increasing production of waste activated sludge (WAS) from wastewater treatment plants presents an inherent environmental burden. In this study, Free nitrous acid combined with potassium ferrate (FNA + PF) pretreatment was used to enhance solubilization and biochemical methane potential of WAS. Results indicated that the maximum removal rates of total suspended solid by PF, FNA, and PF + FNA pretreatment were 21.84%, 38.09%, and 56.17%, respectively. The biochemical methane potential of WAS without pretreatment reached 61.22 L CH4/kg VSS added while this value increased to 147.07 L CH4/kg VSS added after FNA + PF pretreatment (0.06 g/g TSS NaNO2 and 0.25 g/g TSS K2FeO4). Shotgun metagenomic analysis revealed that FNA + PF pretreatment could increase the diversity and stability of microbial communities by shifting methanogenic pathways from strictly acetoclastic to acetoclastic/hydrogenotrophic, thereby enhancing methane production. This study suggested that FNA + PF pretreatment is a promising technology to reduce WAS and enhance methane production by pretreated WAS during anaerobic digestion.

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Due to the large area of agricultural soils contaminated by Cd worldwide, cost-effective and practical method for safety food production are necessary. The roles of micronutrient on reducing Cd accumulation in crops are recently introduced. In the current study, a pot-culture experiment in the greenhouse was conducted to study the foliar spraying of Se (Na2SeO4) and Zn (ZnSO4) on physiological and growth parameters, as well as Cd concentrations in wheat plants grown in Cd-contaminated soil. The foliar was sprayed with four concentration of Se and Zn (0, 10, 20, and 40 mg L-1) at different growth stage (tillering, elongating and heading) and whole wheat plants were collected after maturity. Both foliar spraying with Se and Zn significantly enhanced the photosynthesis, tissue biomass and antioxidant enzyme activity. Additionally, Se and Zn application can also increase Se and Zn concentrations in different plant tissues. Selenium and Zn decreased malondialdehyde (MDA) and Cd concentrations in wheat grains, hulks, leaves, stalks and root in a dose-additive manner. Overall, Se and Zn both efficiently enhanced the wheat growth and Se and Zn concentrations, and simultaneously decreased the Cd concentration in wheat plant. Compared with Zn, Se more efficiently improved wheat growth and reduced Cd concentration in the wheat in a Cd-contaminated soil. Present results suggest that use of foliar spraying, especially Se, could be a cost-effective strategy and could be recommended for remediation of light-or moderate-polluted soils contaminated by Cd.

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A spontaneous development of full-scale anaerobic ammonium oxidation (anammox) process was seldom reported, and its operational parameters could supply references in actual applications. This engineered case indicated that anammox process was suitable for treating relatively high-strength ammonium and organics wastewater due to niche differentiation of biofilm. Results of isotope labelling showed that anammox contributed approximately 40% to N-loss in aerobic unit, but this value increased to 78.3% in anoxic tank. Mass balance showed that N-removal via anammox and denitrification pathways were 38.1 and 23.9 g m-3 d-1, and anammox rate was 1.6 times higher than denitrifiaction. The wild-type anammox granules had a high purity, with anammox accounting for 92.2%. Candidatus Brocadia was the predominant species. Mixing sludge had a higher oxygen tolerance compared with granules, although the latter had a higher anammox activity under anaerobic conditions. Moreover, physicochemical precipitation on the surface of granules may be related to granulation mechanism.

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